Shapiro J A
Department of Biochemistry and Molecular Biology, University of Chicago, Illinois 60637.
J Bacteriol. 1993 May;175(9):2625-31. doi: 10.1128/jb.175.9.2625-2631.1993.
Bacteriophage Mu, one of the best-characterized mobile genetic elements, can be used effectively to answer fundamental questions about the regulation of biochemical machinery for DNA rearrangement. Previous studies of Mu virulence have implicated the Clp protease in repressor inactivation (V. Geuskens, A. Mhammedi-Alaoui, L. Desmet, and A. Toussaint, EMBO J. 13:5121-5127, 1992). These studies were extended by analyzing the phenotypic consequences of clp alleles in two Escherichia coli systems: (i) the periodic replication of Mudlac transposons in colonies and (ii) the action of a Mu prophage in forming araB-lacZ coding sequence fusions. The clpP::CM mutation, which removes the proteolytic subunit of Clp protease, caused a drastic reduction in Mu activity in both systems. The clpA::Tn10 mutation, which removes a regulatory subunit of Clp protease, altered the timing of Mu activity in both systems. A clpA deletion reduced the extent of Mudlac replication in colonies. These results point to temporal changes in Clp proteolysis of the Mucts62 repressor as a key molecular event in the regulation of one class of genomic change in E. coli.
噬菌体Mu是特征最为明确的可移动遗传元件之一,可有效用于解答有关DNA重排生化机制调控的基本问题。先前关于Mu毒力的研究表明Clp蛋白酶参与阻遏物的失活过程(V. 格于斯肯斯、A. 姆哈迈迪 - 阿拉乌伊、L. 德梅特和A. 图桑,《欧洲分子生物学组织杂志》13:5121 - 5127, 1992年)。通过分析clp等位基因在两种大肠杆菌系统中的表型后果,这些研究得到了扩展:(i)Mudlac转座子在菌落中的周期性复制,以及(ii)原噬菌体Mu在形成araB - lacZ编码序列融合中的作用。去除Clp蛋白酶蛋白水解亚基的clpP::CM突变,导致这两种系统中的Mu活性大幅降低。去除Clp蛋白酶调节亚基的clpA::Tn10突变,改变了这两种系统中Mu活性的时间。clpA缺失降低了菌落中Mudlac的复制程度。这些结果表明,Clp蛋白酶对Mucts62阻遏物的蛋白水解作用随时间变化,是大肠杆菌一类基因组变化调控中的关键分子事件。